Screening for DNA Alkylation Mono and Cross-Linked Adducts with a Comprehensive LC-MS3 Adductomic Approach

Alessia Stornetta, Peter W. Villalta, Stephen S. Hecht, Shana J. Sturla, Silvia Balbo

Research output: Contribution to journalArticlepeer-review

33 Scopus citations

Abstract

A high-resolution/accurate-mass DNA adductomic approach was developed to investigate anticipated and unknown DNA adducts induced by DNA alkylating agents in biological samples. Two new features were added to a previously developed approach to significantly broaden its scope, versatility, and selectivity. First, the neutral loss of a base (guanine, adenine, thymine, or cytosine) was added to the original methodology's neutral loss of the 2'-deoxyribose moiety to allow for the detection of all DNA base adducts. Second, targeted detection of anticipated DNA adducts based on the reactivity of the DNA alkylating agent was demonstrated by inclusion of an ion mass list for data dependent triggering of MS2 fragmentation events and subsequent MS3 fragmentation. Additionally, untargeted screening of the samples, based on triggering of an MS2 fragmentation event for the most intense ions of the full scan, was included for detecting unknown DNA adducts. The approach was tested by screening for DNA mono and cross-linked adducts in purified DNA and in DNA extracted from cells treated with PR104A, an experimental DNA alkylating nitrogen mustard prodrug currently under investigation for the treatment of leukemia. The results revealed the ability of this new DNA adductomic approach to detect anticipated and unknown PR104A-induced mono and cross-linked DNA adducts in biological samples. This methodology is expected to be a powerful tool for screening for DNA adducts induced by endogenous or exogenous exposures.

Original languageEnglish (US)
Pages (from-to)11706-11713
Number of pages8
JournalAnalytical Chemistry
Volume87
Issue number23
DOIs
StatePublished - Oct 28 2015

Bibliographical note

Publisher Copyright:
© 2015 American Chemical Society.

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